The novel automated 4-step (anion exchange, desalt, heparin-affinity and size exclusion, in linear sequence) purification protocol for untagged hPCNA results in final purity and yields
of >= 87% and similar to 4 mg L(-1) of original cell culture, respectively, in under 24 h, including all primary sample processing and column equilibration steps. This saves in excess of four full working days when compared to the traditional protocol, producing protein with similar final yield, purity and activity. Furthermore, it limits a time-dependent check details protein aggregation, a problem with the traditional Protocol that results in a loss of final yield. Both automated protocols were developed to use generic commercially available pre-packed columns and automatically prepared minimal buffers, designed to eliminate user and system variations, maximize run reproducibility, standardize yield and purity between batches, increase throughput and reduce user input to a minimum. Both protocols represent robust generic methods for the automated production of untagged hCypA and hPCNA. (C) 2009 JSH-23 Elsevier Inc. All rights reserved.”
“Objective: Optimizing flow and diminishing power loss in the Fontan circuit can improve hemodynamic efficiency, potentially improving
the long-term outcomes. Computerized modeling has predicted improved energetics with a Y-graft Fontan.
Methods: From August to December 2010, 6 consecutive children underwent completion Fontan (n = 3) or Fontan revision (n = 3) using a bifurcated polytetrafluoroethylene Y-graft (18 X 9 X 9 mm in 2, 20 X 10 X 10 mm in 4) connecting the inferior vena cava to the right and left pulmonary arteries with separate graft limbs. The patents underwent magnetic resonance imaging (n = 5) or computed tomography (n = 1). Computational fluid dynamics assessed Fontan hemodynamics, power loss, and inferior vena cava flow splits to the branch pulmonary arteries. The clinical parameters were compared with those from 12
patients immediately preceding the present series who had undergone a lateral Fontan procedure.
Results: Despite longer crossclamp and bypass times (not statistically significant), the Y-graft Fontan patients had postoperative why courses similar to those of the conventional Fontan patients. Other than 2 early readmissions for pleural effusions managed with diuretics, at 6 to 12 months of follow-up (mean, 8 months), all 6 patients had done well. Postoperative flow modeling demonstrated a balanced distribution of inferior vena cava flow to both pulmonary arteries with minimal flow disturbance. Improvements in hemodynamics and efficiency were noted when the Y-graft branches were anastomosed distally and aligned tangentially with the branch pulmonary arteries.